CN103347406A - Methods for improving brain development and cognitive function using beta-hydroxy-beta-methylbutyrate - Google Patents

Abstract

Disclosed are methods for enhancing neural function in an individual, such as an older adult. The methods include administering nutritional compositions comprising HMB to the individual. The nutritional compositions provide benefits for individuals that have or may be at risk of having cognitive decline, cognitive impairment, and neural dysfunction, typically resulting from cognitive diseases associated with neurodegenerative diseases.

Description

Use beta-hydroxy-Beta-methyl butyrate to improve the method for brain growth and cognitive function

Invention field

The disclosure relates to by using institute's choosing method that the nutrition product that comprises beta-hydroxy-Beta-methyl butyrate (HMB) strengthens adult and older adult's neurodevelopment and cognitive function.

Background of invention

Nutrient solution and nutrient powder that the target of making that comprises nutritional labeling is selected are well-known and extensively obtainable, and wherein some can provide unique source of nutrition, and other can provide additional source.These nutriment comprise can water or the powder of other waterborne liquid reconstruct (reconstitue), and i.e. drink (ready to drink) nutrient solution, such as based on the emulsion of breast or albumen or the liquid of non-emulsification.When preparing with selected nutritional labeling, these nutrient solutions are particularly useful.

A kind of such nutritional labeling is beta-hydroxy-Beta-methyl butyrate (HMB).HMB is naturally occurring leucic amino acid metabolite, and leucine becomes known in multiple nutrients product and the replenishers.HMB becomes known for muscle quality and the intensity to help to set up or keep fit in this product in selected individuality.

When being mixed with the oral nutrient product, when wherein product can comprise tablet, capsule, reconfigurable powder (reconstitutable powder), nutrient solution and emulsion, HMB calcium was the common type of HMB.These comprise in the product of HMB some contain other nutrient such as fat, carbohydrate, albumen, vitamin, mineral matter etc.

Recently, there is the interest that increases so-called " intelligence-promoting preparation (smart formulations) " (comprising special nutrition product at brain health and nutrition design) for designing and selling.Many these products are custom-designed at improving cognitive dull-witted and relevant cognitive function decline illness with prevention and disease.Up to now, the success that obtains of these preparations and product is limited.

Therefore, exist for overall improvement is cognitive easily and effectively, and treat the composition of cognitive function decline, cognitive impairment and cognitive illnesses and the needs of method especially.In addition, no matter general health and condition how, if composition and method can be used by extensively individual (and especially older adult), will be favourable so.

Summary of the invention

The disclosure relates to improves cognitive function and/or preventing/treating/the alleviate method of cognitive function decline, cognitive impairment and cognitive illnesses in individual (and especially older adult).Described method comprises the HMB that uses effective dose to individuality.

An embodiment relates to the method for the cognition that improves older adult.Described method comprises composition from the HMB of the amount that comprises that the older adult of effective improvement is cognitive to older adult that use.

Another embodiment relates to the method that is used for the treatment of cognitive illnesses relevant with neurodegenerative disease among the older adult.Described method comprises composition from the HMB of the amount that comprises the older adult's cognitive illnesses of effective treatment to older adult that use.Described neurodegenerative disease can comprise for example Alzheimer disease (Alzheimer's disease), HD (Huntington's disease), Parkinson's (Parkinson's disease), dementia, ALS, apoplexy and schizophrenia.

Another embodiment relates to the method for improving child (toddler), children or teenager's cognitive function.Described method comprises the composition of using the HMB of the amount that comprises effective child of improvement, children or teenager's cognitive function to child, children or teenager.

Method described herein can to individual brain and nerve fiber provides protection and can protect brain and nerve fiber to avoid decline, thereby prevents and/or treats the cognitive illnesses relevant with neurodegenerative disease (particularly in older adult).In addition, using the nutrition product that comprises HMB can be by providing neuroprotective to strengthen nervous function to brain.

Other component of using in alimentation composition and the replenishers can be included with HMB.For example, in one or more embodiments, composition can comprise at least a in albumen, carbohydrate, fat, vitamin and the mineral matter.

Summary of drawings

Fig. 1 is for the figure of the time-histories concentration of HMB in the brain microdialysis liquid of describing rat 1, as assessing among the embodiment 11.

Fig. 2 is for the figure of the time-histories concentration of HMB in the brain microdialysis liquid of describing rat 2, as assessing among the embodiment 11.

Fig. 3 A is for describing the figure of the effect of mTOR phosphorylation state in the Neuro 2A of the HMB cell, as assessing among the embodiment 12.

Fig. 3 B is for describing the figure of the effect of ERK1/2 phosphorylation state in the Neuro 2A of the HMB cell, as assessing among the embodiment 12.

Fig. 3 C is for describing the figure of the effect of Akt phosphorylation state in the Neuro 2A of the HMB cell, as assessing among the embodiment 12.

Fig. 4 is for describing the figure of the effect of albumen synthesis rate in the Neuro 2A of the HMB cell, as assessing among the embodiment 12.

Fig. 5 A-5D is described under the situation that different inhibitor do not exist or exist the figure of the effect of MEF2 transcription factor expression in the Neuro 2A of the HMB cell, as assessing among the embodiment 12.

Fig. 6 is for the figure of the effect of the propagation of describing the Neuro 2A of HMB cell and viability, as assessing among the embodiment 12.

Fig. 7 is the figure that describes in the forced swimming test as pass through the antidepressant effect of measured HMB in motionless incubation period, as assessing among the embodiment 13.

Detailed Description Of The Invention

Method of the present disclosure relates to the method (especially in older adult) of utilizing the composition that contains HMB to improve cognitive function and alleviating cognitive function decline, cognitive impairment and cognitive illnesses.Hereinafter describe these and other basic or optional element or restriction of method of the present disclosure in detail.

Unless stipulate that in addition term " older adult " refers at least 45 years old ages, comprises at least 50 years old ages, comprises at least 55 years old ages, comprises at least 60 years old ages, comprises at least 65 years old ages, comprises at least 70 years old ages, comprises at least 75 years old ages, comprises at least 80 years old ages or comprises about 45 years old of age to about 80 years old of the age more greatly and also, further comprises that about 55 years old of age is to about 80 years old individuality of age as used herein.

Unless stipulate that in addition term " HMB calcium " refers to the calcium salt of beta-hydroxy-Beta-methyl butyrate (being also referred to as beta-hydroxy-methylbutanoic acid, beta-hydroxy isovaleric acid or HMB) as used herein, it is the form of monohydrate the most normally.Unless stipulate in addition, all wt, percentage and the concentration that are used for sign HMB calcium as used in this article are based on the weight of HMB calcium monohydrate.

Unless stipulate that in addition term " spray-dried powders " refers to nutrient powder as used herein, wherein most of components (comprising HMB) are during manufacture through homogenizing and carrying out spray-drying process subsequently.Other composition can be added in the spray-dried powders by dry type fusion (dryblending), as long as HMB had before homogenized and spray-drying at least.

Unless other regulation, term " fat " and " oil " exchange and are used in reference to the lipid matter that derives from plant or animal or process from plant or animal as used herein.These terms also comprise the synthetic fat metallic substance, as long as this synthetic is suitable for being administered orally in the people.

Unless other regulation, term " stable storing " refers to packaged and then at 18-24 as used herein ^oC stores the commercial stable nutrient solution of back maintenance at least 3 months (comprise about 6 months to about 24 months, and also comprise about 12 months to about 18 months).

Unless stipulate in addition, term " nervous function " refers to that brain and neuronal tissue support and keep the function of cognitive ability (for example learning and memory) as used herein.By contrast, unless stipulate in addition, term " neurological dysfunction " refers to that memory or cognitive function reduce as used herein.For example, the ability reduction of learning ability or memory information is regarded as neurological dysfunction.In some embodiments, neurological dysfunction can be the result of aging or neurodegenerative disease.

Unless stipulate in addition, term " neuroprotective " refers to protect existing neuron and nerve fiber to avoid Apoptosis or sex change as used herein; The existing neuron of protection and nerve fiber antagonism physical damnification; Regenerate with stimulating neuronal.

Unless stipulate that in addition term " cognition " refers to participate in the mental process of acquire knowledge and understanding as used herein, comprise that thinking, cognition, memory, judgement and problem solve." cognition " comprises the high-level function of brain and contains language, the imagination, perception and plan.

Unless stipulate in addition, term " alimentation composition " or " nutrition product " refer to nutrient solution and nutrient powder as used herein, its latter can be formed nutrient solution by reconstruct, all these all comprise in HMB and fat, albumen and the carbohydrate one or more and to be suitable for human oral edible.

Unless stipulate that in addition term " nutrient solution " refers to be the nutrition product of namely drinking liquid form as used herein, and refers to by the prepared nutrient solution of the nutrient powder described herein of reconstruct before use.

Unless stipulate in addition, term " Cheng Qing liquid basically " refers to be substantially free of the nutrient solution of fat as used herein; That is, add to help to make the fat of liquid except the intrinsic fat of raw material or with low concentration, described liquid does not contain the fat of interpolation.In this context, term " not fatty " means that in the weight of nutrient solution, described liquid generally contains less than 1.0 %, more is generally less than 0.5 %, and more is generally less than 0.1 %, comprises the fat of 0 %.These nutrient solutions of clarifying basically are can flow or drinkable liquid at about 1 to about 25 ℃.

Unless other regulation, all percentages, umber and ratio are all in the weight of total composition as used herein.Unless stipulate that in addition all this weight are based on activity level when it relates to ingredients listed, therefore do not comprise to be included in solvent or the accessory substance that is purchased in the material.

Unless regulation or mentioned context hint clearly that on the contrary all should comprise corresponding plural feature or restriction to mentioning of odd number feature of the present disclosure or restriction in addition, vice versa.

Unless the context of stipulating in addition or mentioning combination is clear and definite hint on the contrary, all combinations of method or process steps all can any order be carried out as used herein.

The various embodiments of alimentation composition of the present disclosure also can be substantially free of any basis or feature optional or that select described herein, and condition is that remaining nutrition product still contains all required composition or features as described herein.Unless stipulate that in this context and in addition term " is substantially free of " and means by weight, selected nutrition product contains the optional member less than function, be generally less than 1%, comprise less than 0.5%, comprise less than 0.1%, and also comprise 0% basis this optional or that select.

Alimentation composition described herein and method can comprise the fundamental of composition and method as described herein and this paper describes or be used in addition that nutrition product uses any other or optional key element are made up of it, or are made up of it basically.

Product form

It is can be any known or be applicable to the product form preparation of oral or parenteral administration in addition to can be used for the composition that comprises HMB in the method for the present disclosure.The Orally taken product form is normally preferred, and comprises that any solid, liquid or the powder formulation that is applicable to this paper purposes, condition are that this preparation allows oral delivery securely and effectively from basis and other selected composition of selected product form.

The limiting examples that is applicable to the solid nutrition product form of this paper purposes comprise snacks and generation the meal product, comprise being mixed with following those: rod, bar, biscuit or bread or cake or other baked goods, frozen liq, candy, breakfast cereal preparation, powder or granular solids or other particle, snack chip or sting sheet, freezing or boiling staple food (frozen or retorted entrees) etc.

The limiting examples that is applicable to the fluid product form of this paper purposes comprise snacks and generation meal product, hot drink or cold drink, carbonic acid or noncarbonated beverage products, fruit juice or other acidifying drinks, the beverage based on breast or soybean, bland (shakes), coffee, tea, nutrition composition etc.The most common suspension or the emulsion of being mixed with of these fluid compositions, but also can any form that other is fit to prepare, such as the liquid of clarification, the liquid of clarifying basically, solution, liquid gel etc.

Other limiting examples of the Orally taken product form that is fit to comprises semisolid or semi-liquid composition (for example pudding, gel) and more conventional products forms (such as capsule, tablet, Caplet, pill etc.).Provide the amount of composition of the HMB of effective dose can be contained in one or more independent formulations (for example can a tablet or a plurality of tablet that every day, single or multiple dosage were used) for the target user.

For product form (such as lozenge, tablet (for example chewable tablets, coated tablet etc.), paste or gel), weight in product form, the concentration range that the amino acid admixture can be prepared be generally most about 5% to about 50%, comprise about 15% to about 33%, and also comprise about 15% to about 25%, its all can with excipient or other composition (such as carbohydrate, acidulant, flavor enhancement and colouring agent) combination.

The composition that comprises HMB can be prepared with the nutrient of enough kinds and amount, so that single, main or extra-nutrition source to be provided, or is provided for the special nutrition product suffering from the individual of specified disease or illness or have the target nutritional benefits.

Beta-hydroxy-Beta-methyl butyrate (HMB)

The composition that is used for method of the present disclosure comprises HMB, and it means that described composition is by the most common form with the calcium monohydrate of interpolation HMB() prepare or otherwise prepare in final products, to contain calcium and HMB.Any source of HMB all is applicable to this paper, and condition is that final products contain HMB, although this source is preferably HMB calcium, and adds in the described composition by original state in process for preparation the most usually.

Term " the HMB calcium of interpolation " refers to the calcium salt of HMB as used herein, the form of the most common calcium salt monohydrate with HMB, and it is as the HMB source that is added in the described nutrition product.

Although HMB calcium monohydrate is the preferred HMB source for this paper purposes, but other suitable source can comprise free acid, salt, anhydrous salt, ester, lactone form or the HMB of other products form of the HMB of biological utilisation form is provided from described nutrition product in addition.The non-limiting example of suitable salt that is used for the HMB of this paper purposes comprises sodium salt, sylvite, magnesium salts, chromic salts, calcium salt or other the nontoxic salt form of hydration or anhydrous HMB.HMB calcium monohydrate is preferred, and commercially available from Salt Lake City, the Technical Sourcing International (TSI) of Utah.

Be applicable to the HMB range of concentrations in the nutrient liquor composition of using in the described method, weight in nutrient liquor composition, can be for being up to 10%, comprise about 0.01% to 10% and also comprise about 0.1% to about 5.0% and also comprise about 0.5% to about 2.0% and also comprise about 0.4% to about 1.5%.In a particular, in the weight of nutrient liquor composition, HMB is present in the nutrient liquor composition with about 0.67% amount.

Be applicable to the HMB range of concentrations in the nutritive solid composition that uses in the described method, weight in the nutritive solid composition, can be for being up to 10%, comprise about 0.01% to 10% and also comprise about 0.1% to about 7.0% and also comprise about 1.0% to about 5.0% and also comprise about 1.0% to about 4.0%.In a particular, in the weight of nutritive solid composition, HMB is present in the nutritive solid with about 3.2% amount.

Describe the alimentation composition of using to individuality as this paper about 0.1 HMB to about 10 grams/sky can be provided, comprise about 0.1 HMB to about 5.0 grams/sky.Correspondingly, alimentation composition is edible at every turn can to provide about 0.5 to about 2.5 to restrain, comprise that about 1.0 to about 1.7 restrain, comprise the HMB of about 1.5 grams, and wherein once edible can be the nutritive solid of about 240ml instant nutrient liquid or about 240ml reconstruct.In a particular, HMB provides with the level of about 1.58 gram/every 240ml.It is edible or more times is edible to use once a day edible or every day four times edible, every day three times twice of edible, every day to individuality, to accept the HMB of aequum from alimentation composition.

Macronutrient

The composition that comprises HMB that is applicable to described method as disclosed herein can further comprise one or more other macronutrients except all comprising as described herein HMB, comprise adipose-derived, carbohydrate source and dietary protein origin.

Optional macronutrient with other must or the adding ingredient combination can provide each edible or every dosage the most nearly 1000 kcal energy, comprise each edible or about 25 kcal of every dosage to about 900 kcal, also comprise about 75 kcal to about 700 kcal, also comprise about 100 kcal to about 500 kcal, also comprise about 150 kcal to about 400 kcal and comprise that also about 200 kcal are to the energy of about 300 kcal (optimum as single, portioning eats or dosage).

Albumen, liquid and the carbohydrate of a lot of separate sources and type are known, and can be used for the composition that comprises HMB described herein, condition be selected nutrient for Orally administered be safe and efficient, and with must composition and the composition of other interpolation be compatible.

Be applicable to that the carbohydrate in the composition can be simple carbohydrates, complex carbohydrate or its variant or combination.The limiting examples of suitable carbohydrate comprises hydrolysis or converted starch or cornstarch, maltodextrin, glucose polymer, sucrose, corn syrup, corn-syrup solids, the carbohydrate that derives from rice, glucose, fructose, lactose, high-fructose corn syrup, indigestible compound sugar (for example FOS), solvable or soluble fiber, honey, sugar alcohol (for example maltitol, antierythrite, D-sorbite) and combination thereof.

Except the HMB component as described herein, be applicable to that albumen in the composition comprises albumen or the dietary protein origin of hydrolysis, partial hydrolysis or non-hydrolysis, and can derive from any known or other suitable source, such as breast (for example casein, whey), animal (for example meat, fish), cereal (for example rice, corn), plant (for example soybean) or its combination.

Be applicable to that the fat in the composition comprises coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, high oleic safflower oil, MCT oil (medium chain triglyceride), sunflower oil, high oleic sunflower oil, palm and palm-kernel oil, palm olein, Canola oil, marine facies oil, cottonseed oil and combination thereof.

The concentration of the fat in composition of the present disclosure, albumen and carbohydrate or amount can have remarkable change according to specific product form (for example, solid, liquid, powder) and various other preparation and target dietary requirements.Preparation in the most common any calorie of scope of in following table, describing (embodiment A-D) of these macronutrients.

Optional member

The alimentation composition that comprises HMB can further comprise other optional member, and it can change the physics, nutrition, chemistry of product, joyful or processing characteristics maybe serves as medicine or extra nutrition composition when being used for target group.Many this optional members are known or are suitable in addition in other alimentation composition, and also can be used for alimentation composition described herein, condition is that this optional member is safe and efficient and compatible with other composition with the basis in the selected product form for dosage forms for oral administration.

The limiting examples of this optional member comprises anticorrisive agent, antioxidant, emulsifying agent, buffer, FOS, chromium picolinate, pharmaceutically active agents, other nutrient, colouring agent, flavor enhancement, thickener and stabilizing agent etc. as described herein.

Composition can further comprise vitamin or relevant nutrient, and its limiting examples comprises vitamin A, vitamin D, vitamin E, vitamin K, thiamine, riboflavin, Cobastab ₆, Cobastab ₁₂, carotenoid, nicotinic acid, folic acid, pantothenic acid, biotin, vitamin C, choline, inositol, its salt and derivative, and combination.

Described composition can further comprise mineral matter, and its limiting examples comprises phosphorus, magnesium, calcium, iron, zinc, manganese, copper, sodium, potassium, molybdenum, chromium, selenium, chloride and combination thereof.

Described composition can also comprise one or more flavor enhancements or screening agent.Suitable flavor enhancement or screening agent comprise natural and artificial sweetener, sodium source such as sodium chloride and hydrocolloid such as guar gum, xanthans, carrageenan, gellan gum, gum arabic and combination thereof.

Manufacture method

The nutrient liquor composition that contains HMB can be produced by the method for any preparation nutrient solution (comprising that emulsion is such as the nutritional emulsions based on breast) known or that be fit in other side.

In a kind of suitable manufacture method, use following preparation nutrient solution: at least three kinds of independent slurries comprise fat bag albumen (protein-in-fat (PIF)) slurry, carbohydrate-mineral matter (CHO-MIN) slurry and water bag albumen (protein-in-water (PIW)) slurry.The PIF slurry is by heating and mix selected oil (for example Canola oil, corn wet goods), forms continuing to add under heating and the stirring condition emulsifying agent (for example lecithin), liposoluble vitamin and a part of total protein (for example lactoprotein concentrate etc.) subsequently.The CHO-MIN slurry forms by under condition of heating and stirring following material being added in the water: mineral matter (for example potassium citrate, dipotassium hydrogen phosphate, natrium citricum etc.), trace and ultra-trace mineral (TM/UTM pre-composition), thickener or suspending agent (for example Avicel (Avicel), gellan (gellan), carrageenan) and HMB.The CHO-MIN slurry that obtains kept 10 minutes under lasting heating and stirring condition, added other mineral matter (for example potassium chloride, magnesium carbonate, KI etc.) and/or carbohydrate (for example FOS, sucrose, corn syrup etc.) subsequently.The PIW slurry forms by under heating and stirring condition residual protein (for example casein sodium, soybean protein concentrate etc.) being mixed in the water subsequently.

Subsequently the slurry that obtains is blended together under condition of heating and stirring and pH is adjusted to required scope, be generally 6.6-7.0, subsequently composition is carried out high temperature, short time (HTST) processing, during this period with composition heat treatment, emulsification with homogenize, make its cooling subsequently.Add water soluble vitamin and ascorbic acid, in case of necessity pH is adjusted to required scope again, add flavor enhancement, and add water to reach required total solid level.Subsequently composition sterile is packed to form the aseptic packaging nutritional emulsions, or add composition to the stable container of boiling and stand boiling sterilization subsequently to form the nutritional emulsions of boiling sterilization.

Under the situation that does not break away from spirit and scope of the present disclosure, the manufacture method of nutritional emulsions can be carried out with those methods in addition described herein.Therefore, it is illustrative rather than restrictive in all fields that the present embodiment is considered to, and all changes and equivalents are also in description scope of the present disclosure.

This nutritive solid such as the spray-drying nutrient powder, can prepare by any set known or other otherwise effective technique that is applicable to preparation and preparation spray-drying nutrient powder.

This spray-drying step can comprise any spray drying technology known or that be applicable to the production nutrient powder in addition equally.Many different spray drying process and technology become known for nutrition field, and it all is applicable in the manufacturing of spray-drying nutrient powder of this paper.

A kind of method for preparing spray-dired nutrient powder comprise form and homogenize the water paste that comprises HMB and optional albumen, carbohydrate and fat or liquid and subsequently with described slurry or liquid spray-drying to produce spray-dired nutrient powder.Described method may further include that spray-drying, dry type are mixed or otherwise adds the step of other nutritional labeling (comprise in the composition described herein any or multiple) in the spray-dired nutrient powder.

This manufacture method preferably uses HMB calcium (it is the most common with the preparation of HMB calcium monohydrate) to originate to prepare as the HMB that uses in the described method.

Using method

The composition that comprises HMB as described herein especially can be used cognitive or treatment specific knowledge disease or illness (such as cognitive illnesses relevant with neurodegenerative disease among the older adult) with overall improvement to individual (generally including adult, older adult, child, children and teenager).Individuality generally can be healthy maybe can suffer from the decline of (or being in the risk of suffering from) cognitive function, cognitive impairment (comprising mild cognitive impairment (MCI)), be losing one's memory, generally recall problem, cognitive illness or neurodegenerative disease (such as Alzheimer disease, HD, Parkinson's, dementia, ALS, apoplexy and schizophrenia).Specifically for child, children and teenager, the composition that contains HMB can improve overall cognitive function and brain growth.

The nervous function that the alimentation composition that contains HMB strengthens is individual (comprising the older adult that can significantly be benefited from the nervous function that strengthens, because nervous function generally reduces with age growth).Particularly, the older adult who suffers from neurological dysfunction especially can benefit from the composition that contains HMB.

In specific embodiments, the individuality of accepting to contain the composition of HMB can be to suffer from cognitive illness or be in the risk that develops into cognitive illness or be in older adult in the risk that develops into cognitive function decline or cognitive impairment (comprising MCI).Be in the individuality that develops in cognitive function decline or cognitive illness or damaged " risk " comprise have the specific gene form that is called APOE-e4, suffer from hypertension, diabetes, depression, hypertension, cholesterol raise individual and/or individuality with family history of the cognitive function decline, cognitive impairment and/or the cognitive condition/disease that cause because of the age.

The method described herein that utilization contains the composition of HMB further relate to after using this composition (the most common about 1 month to about 10 years, comprised about 1 month to about 1 year and further comprised after used every day between about 6 months extended period in about 1 month) to individuality provide following one or more: (1) is supported and is kept and be grown up and older adult's nervous function; (2) enhancing adult and older adult's nervous function; (3) to adult and older adult's neuroprotective; (4) cognitive illnesses relevant with neurodegenerative disease among preventing/treating adult and the older adult; (5) cognitive function decline or the cognitive impairment among preventing/treating adult and the older adult; (6) improve child, children and teen-age cognitive ability.In one embodiment, use the composition that contains HMB every day and reached at least 1 year during.

Nutrition product as required dosage forms for oral administration so that the desired nutritional level to be provided, the most common with once a day to twice edible form, with once a day or the form of twice or more times broken dose, for example edible deal scope is generally about 100 to about 300 ml, comprise about 150 to about 250 ml, and comprise that about 190 ml are to about 240 ml.

Embodiment

Following examples have illustrated the alimentation composition of the HMB of containing of the present disclosure and particular and or the feature of method.Described embodiment only provides and should not be construed as the restriction disclosure for purposes of illustration, because may there be many variations that do not deviate from spirit and scope of the present disclosure in it.Unless stipulate that in addition the amount of all embodiment is the percentage by weight based on the gross weight of composition.

The composition that contains HMB of embodiment is according to well-known for the preparation of the prepared nutrition product of the manufacture method of nutritional emulsions and spray-dired nutrient powder in the nutraceutical industry.

Embodiment 1-5

Embodiment 1-5 explanation is applicable to the nutrient powder of the spray-dired HMB of containing in the disclosure method, during its composition is listed in the table below.With separately batch preparation, and water reconstruct before use is to required target component concentration by spray drying process for these products.Unless stipulate in addition, the amount of all the components goes out with the kilogram ordered series of numbers of per 1000 kg batch products.

Embodiment 6-10

Embodiment 6-10 illustrates the nutritional emulsions embodiment that contains HMB of the present disclosure, during its composition is listed in the table below.Unless stipulate in addition, all amounts go out with kilogram ordered series of numbers of per 1000 kg batch products.

Embodiment 11

In the present embodiment, after dosage forms for oral administration HMB, analyze appearance and its concentration of passing in time of HMB in the brain interstitial fluid.

At first, with the Sprague-Dawley rat of heavy 400-500 g (Charles River, France) in the cage of constant room temperature (22 ± 2 ℃) and 45-55% humidity according to regular 12 little time/secretly timetable stable breeding.Food and water can arbitrarily obtain.(RD 2101-2005, carry out about animal used as test nursing and the state's laws that use and EC policy by system criterion 86/609/CEE) according to meeting for the program that relates to animal and its nursing.

Conduit inserts in the hippocampus of each anesthetized rat with ground, the location of the coordinate brain domain described in the stereotaxic atlas of rat.Allow rat from surgery recovery at least 3 days.First day of experiment, brain microdialysis probe (the 4mm film, from BASi, West Lafayette, Indiana obtains for MD-2204, BR-4) is inserted in the conduit of each rat.(aCFS Harvard apparatus #597316) pours into probe with the constant rate of speed of 2 μ l/min with Ringer's mixture.Rat is retained in the cage with free-moving relatively space.Automatically collected the dislysate sample every 20 minutes.Give heavy dose of HMB (250mg/kg body weight) and continue another 160-220 min of sampling by gavage at experiment beginning back 100 min.

Use UPLC-MS to measure HMB.Particularly, by 0.2 μ m NF microfiltration, usefulness the water gaging of etc.ing dilutes with the sample of 30-40 μ l among the aCSF, and injects subsequently among the UPLC-TQD (Acuity-TQD system, from Waters Corporation, Milford, Massachusetts acquisition).Use BEH Hillic post 1.7 μ m; 2.1x150 mm, use are by H ₂The gradient analysis HMB that O:MeCN 0.1% formic acid is formed.Mass spectrograph is set at ion mode ES+.

Analyzed two rats at different days.Give tube feed the 5th grade of timesharing.As shown in fig. 1, use and occurred HMB (the 7th fraction) in rat 1 brain in back 40 minutes, and sharply increase subsequently.Gather fraction, behind tube feed above 2.5 hours.At this moment, the HMB concentration level does not also reduce.The Cmax of measured HMB is 10.5 ppm.

Fig. 2 shows the result of rat 2.Specifically, the fraction collection continues to reduce to detect HMB concentration between more long-term.The time-histories concentration of HMB in the brain microdialysis liquid of rat 2 is followed and rat 1 similar pattern.It detects for the first time in the 7th fraction and having to go to the toilet in back 2 hours 20 minutes increases severely adds (12-13 fraction) using, and wherein reaches Cmax (greater than 4 ppm).Thereafter the HMB concentration level reduces, although (surpass 3.5 h behind the tube feed) when experiment finishes, it does not reach foundation level as yet.

According to described result, the HMB of dosage forms for oral administration appears in the brain interstitial fluid, though it means that HMB has relative water-wet behavior, it still can pass the blood-brain barrier in the rat microdialysis experimental model.

Embodiment 12

In the present embodiment, analyze the effect that signal synthetic relevant with cell proliferation with albumen in the nerve cell of HMB (Neuro 2A) transmits the adjusting of cascade.Particularly, analyze HMB and induced neuronal development and plasticity and the effect of giving neuroprotective as nutrition composition.

Neuronal development and cynapse transmission are to be subjected to the plastic process that the condition change influences in extracellular and the cell.Protein kinase is most important to the adjusting of these phenomenons, and wherein several (for example mitogen activated protein kinase (ERK), phosphoinositide-3 ' kinases/AKT (PI3K/AKT) and calcium/calmodulin-dependent kinases (CaMK)) have the effect of generally acknowledging in neuronal development and synaptic plasticity.Recently, the mammal target (mTOR) for serine/threonine protein kitase, rapamycin has had increasing concern.In fact, mTOR as the adjusting of the different phase of neuronal development and axon guidance, dendritic spines form take place and the several forms of long-term synaptic plasticity in have crucial effects.

Myocyte enhancer factor 2 (MEF2) also plays an important role in neuronal survival.The approach of regulating the activity of MEF2 in neuronal survival comprises hormone/factor or the post-stimulatory p38 MAPK of film depolarising (ERK) and PI3-AKT phosphorylation.In addition, in neurodegenerative disease, the huge autophagy of inhibition induced strong of MEF2 degraded or mTOR approach causes neuronal death.Although it is unknown that mTOR regulates the mechanism of MEF2, several observed results point out, the signal transmission of the combination by mTORC2 and AKT/PKB may need for the neuronal survival mechanism that MEF2 triggers.

For the transfection experiment in the present embodiment, the Neuro-2A cell that the 80-90% of use growth in the Eagle's medium (DMEM) of DulbeccoShi improvement converges.Cellular exposure is reached 5 hours in plasmid pGL3-4xMEF, and described plasmid pGL3-4xMEF contained MEF2D in conjunction with 4 tandem copies of dna sequence dna before the dna sequence dna of coding firefly luciferase.Effector is added into reaches 18 h in the culture medium and measure uciferase activity.In order to study the inhibition of signal transmission factor, the inhibitor that phosphatidyl-inositol 3 kinases (LY294002), ERK 1/2 (PD98059) and mTORC1 is activated (rapamycin) is added in the cell.

By measuring the effect of incorporating to measure albumen synthesis rate in the Neuro-2A cell of HMB into of [3H]-tyrosine in the cell protein storehouse.Measure the phosphorylation state of mTOR and rise/down-regulation protein kinases (Akt, ERK 1/2) by Western blotting.Cell proliferation/viability is measured with MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide) colorimetric estimation.

Data are represented with mean value ± SEM.Use Student t-to check analysis result.

The effect of the phosphorylation of mTOR and upstream regulation in the Neuro 2A of Fig. 3 A-3C demonstration HMB cell.Under above-mentioned cell culture condition, HMB produces the remarkable increase (Fig. 3 A) of mTOR phosphorylation.In addition, HMB has strengthened the phosphorylation state of Akt in this clone (Fig. 3 C) and ERK 1/2 (Fig. 3 B), and it is two kinds of main upstream regulation of mTOR signal pipeline in the mammalian cell.In addition, the HMB higher mTOR activation of inducing promotes that albumen is synthetic increases that (20%, HMB is than contrast, Fig. 4).

Fig. 5 A shows the effect of the expression of MEF2 transcription factor among the Neuro2A of HMB.Produce the remarkable increase of MEF2 dependent transcription uciferase activity with the HMB supplementing culture medium.LY294002, PD98059 and rapamycin inhibitor also are shown among Fig. 5 B-5D the effect of MEF2 transcription factor.LY294002 or PD98059 block the increase of the MEF2 of HMB mediation fully.Yet the MEF2 that rapamycin is induced HMB expresses not influence.LY294002 and PD98059 work to block expression in the upstream of mTOR.It suppresses the mTOR activity of most of (if not all).By contrast, influence to the rapamycin allosteric mTOR substrate interaction and only optionally block the subclass (by those compound-mediated activity of mTORCl, rather than by other activity of mTORC2 mediation) of mTOR activity.This is consistent with following discovery: rapamycin does not hinder neuronal survival, dendron growth and complexity or the action potential of PI3/AKT activation target.Above-mentioned data show that Ras/ERK and PI3/AKT approach may all need for the MEF2 activity that HMB mediates, thereby with downstream effect of MEF2 as Ras/ERK and PI3/AKT approach.

Fig. 6 shows the effect of the neuron propagation of HMB and viability.As shown in Figure 6, HMB stimulating neuronal propagation and viability.

The data of present embodiment (using Neuro 2A cell) demonstration HMB serves as can regulate the key signal pipeline as the effector of the mTOR-raptor/rictor-MEF2 of adjusting key foundation cell processes (the synthetic and propagation such as albumen).Correspondingly, HMB can be by regulating the control that neuron signal pipeline helps nervous system development and neurodegenerative disease.

Embodiment 13

In the present embodiment, analyze the effect of the rat anxiety of HMB/depressed behavior.

The depressed forced swimming test (FST) that present embodiment was described based on people such as Porsolt in 1978.For begin the experiment, with rat individually place contain 25 ℃ of water of 30cm upright cylinder (highly: 40 cm; Diameter: reach 15 minutes 21.5 cm) (i.e. " pre-swimming ").After this pre-swimming phase, shift out rat and allow it dry in the outer cover of heating, return then in its cage.Rat oral is used the dosage of the HMB of 250 mg/kg body weight or 500 mg/kg body weight subsequently.

After 24 hours, make rat accept test swimming, wherein rat is placed again cylinder to reach 5 minutes and assess motionless and total duration escape behavior.Test swimming is recorded a video and assessed motionless incubation period subsequently, and it is not for observing the point that mobile and rat is in " timber " state (i.e. head upwards, under the Caudad, under foreleg slight bending and the back leg sensing).Test swimming preceding 4 hours and 1 hour, rat oral was used its dosage separately of the HMB of 250 mg/kg body weight or 500 mg/kg body weight.

As shown in Figure 7, more the HMB of high dose prolongs motionless incubation period (P=0.07).These data show that using HMB has the effect of antidepression sample.

Claims (15)

1. be used for the method for the older adult's of improvement cognition, described method comprises composition from the beta-hydroxy-Beta-methyl butyrate of the amount of the cognition that comprises the described older adult of effective improvement to described older adult that use.

2. the process of claim 1 wherein and use beta-hydroxy-Beta-methyl butyrate to described older per day for adults.

3. the method for claim 2 is wherein used beta-hydroxy-Beta-methyl butyrate of about 0.1 g/ days to about 10g/ days to described older adult.

4. the method for claim 2 is wherein used beta-hydroxy-Beta-methyl butyrate of about 0.1 g/ days to about 5.0 g/ days to described older adult.

5. the process of claim 1 wherein that described composition is the following form that is selected from: nutrient powder, nutritional emulsions and supernatant liquid.

6. the process of claim 1 wherein that described older adult suffers from the cognitive function decline.

7. the process of claim 1 wherein that described older adult suffers from neurological dysfunction.

8. the process of claim 1 wherein use described beta-hydroxy-Beta-methyl butyrate reach at least 1 year during.

9. the method that is used for the treatment of cognitive illnesses relevant with neurodegenerative disease among the older adult, described method comprise composition from the beta-hydroxy-Beta-methyl butyrate of the amount that comprises the cognitive illnesses among the described older adult of effective treatment to described older adult that use.

10. the method for claim 9, wherein said neurodegenerative disease is selected from Alzheimer disease, HD and Parkinson's, dementia, ALS, apoplexy and schizophrenia.

11. the method for claim 9 is wherein used beta-hydroxy-Beta-methyl butyrate to described older per day for adults.

12. the method for claim 11 is wherein used beta-hydroxy-Beta-methyl butyrate of about 0.1 g/ days to about 10g/ days to described older adult.

13. the method for claim 11 is wherein used beta-hydroxy-Beta-methyl butyrate of about 0.1 g/ days to about 5.0 g/ days to described older adult.

14. the method for claim 9, wherein said composition are the following form that is selected from: nutrient powder, nutritional emulsions and supernatant liquid.

15. be used for improving the method for child, children or teen-age cognitive function, described method comprises the composition of using the beta-hydroxy-Beta-methyl butyrate of the amount that comprises the described child of effective improvement, children or teen-age cognitive function to described child, children or teenager.

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